title: [Study on no-fusion expression and purification of Glycine max dehydration-responsive element-binding protein (GmDREB1) expressed in E. coli].
abstract: OBJECTIVE
  To obtain enough GmDREB1 protein comparable with native protein for further safety assessment of the protein.
  
  
  METHODS
  The GmDREB1 gene was cloned into no-fusion expression vector pBV220 and the recombinant vector pBV220-GmDREB1 was obtained. The constructed vector was transformed into expression host E. coli DH5a. The protein expression was optimized by improving the codon, induced expression conditions and selecting the appropriate vector. The protein was obtained by cation exchange chromatography and gel filtration chromatography and identified by N-terminal amino acid sequencing, western blotting and activity determination.
  
  
  RESULTS
  The soluble protein was expressed efficiently in E. coli DH5a containing the optimized target gene by 42 degrees C induction for 3 hours and the purified protein consistent with the native protein was obtained through the chromatography.
  
  
  CONCLUSIONS
  The results of this study illustrated that the GmDREB1 protein could be acquired through prokaryotic host expression which had comparable N-terminal amino acid sequences, immunogenicity and biological activities with those of native GmDREB1 protein.
authors.affiliation: Institute for Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing, China.
authors.forename: Zhaolong
authors.initials: Z
authors.lastname: Gong
authors.forename: Qin
authors.initials: Q
authors.lastname: Zhuo
authors.forename: Jinghuan
authors.initials: J
authors.lastname: Wu
authors.forename: Jianhua
authors.initials: J
authors.lastname: Piao
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Yang
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D013025:Glycine max / Q000378:metabolism*; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D010940:Plant Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors